Development of multivalent, ultrapotent nanobody cocktails for SARS-CoV-2 neutralization

开发用于中和 SARS-CoV-2 的多价、超强纳米抗体混合物

• 批准号: 10444442
• 负责人: William Paul Duprex
• 金额: $ 76.9万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-09 至 2022-02-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10444442
• 关键词: 2019-nCoV; Aerosols; Affinity; Aftercare; Antibodies; Binding; Biological Availability; Biomedical Engineering; COVID-19; COVID-19 outbreak; COVID-19 therapeutics; COVID-19 treatment; Cells; Clinical; Clinical Research; Clinical Trials; Communities; Computer software; Coronavirus; Deposition; Development; Diagnostic Reagent; Dose; Drug Storage; Epitope Mapping; Epitopes; Event; Future; Genotype; Goals; Hamsters; Human; Immunoglobulin Fragments; Immunoglobulin G; Infection; Inhalation; Innovative Therapy; Lung; Microbe; Modeling; Mutation; Nebulizer; Pharmaceutical Preparations; Primates; Production; Property; Proteomics; Pulmonary Pathology; Reporting; Research; Resistance; Resolution; Respiratory System; SARS-CoV-2 infection; SARS-CoV-2 variant; Science; Structure; System; Technology; Therapeutic; Therapeutic Agents; Translations; Viral; Viral Antibodies; Viral Load result; Viral Pneumonia; Virus; aerosolized; base; clinical application; cost; cost effective; design; drug development; drug quality; efficacy evaluation; emerging pathogen; flexibility; global health; improved; in vivo; insight; interdisciplinary approach; interest; multidisciplinary; nanobodies; novel; novel therapeutics; novel vaccines; pandemic disease; pathogen; preclinical efficacy; preclinical evaluation; prevent; protein folding; rational design; receptor binding; respiratory virus; structural biology; therapeutic candidate; therapeutically effective; translational potential; variants of concern

PROJECT SUMMARY/ABSTRACT The outbreak of COVID-19 has severely impacted global health and the economy. Cost-effective, highly efficacious therapeutics are urgently needed. Camelid VHH antibodies or nanobodies (Nbs) are small, highly stable, easily bioengineered, and can be rapidly and economically manufactured from microbes. They are highly robust and are flexible for administration, including possible delivery by nebulization. Together these unique properties of Nbs make their uses against respiratory viruses such as SARS-CoV-2 especially appealing. We recently developed a disruptive proteomic technology for large-scale identification of multi-epitope, drug- quality Nbs (Xiang et. al, Cell Systems. 2021). Using this technology, we identified > 8,000 high-affinity Nbs for the SARS-CoV-2 spike (S) receptor-binding domain (RBD) including Nbs that target highly neutralizing epitopes with sub-pM affinities and can neutralize SARS-CoV-2 at sub-ng/ml concentrations, which are unprecedented for antiviral antibody fragments. Structural proteomics revealed multiple distinct epitopes and potential neutralization mechanisms. Bioengineering of multi-epitope and multivalent constructs improved the potency to below 0.1 ng/ml (Xiang, et. al, Science. 2020). Most recently, we have demonstrated the high preclinical efficacy of an ultrapotent and stable trimeric Nb construct (PiN-21) for inhalation treatment of SARS- CoV-2 infection in a sensitive COVID-19 model (Nambulli, et. al, Science Advances. 2021). Intranasal delivery of PiN-21 at 0.6 mg/kg substantially reduces viral burdens in both airways. Critically, aerosol delivery of PiN-21 at 0.2 mg/kg decreases lung viral titers by 6-logs, minimizing lung pathology post-infection and preventing viral pneumonia. Combined with the marked stability and low production cost, this innovative therapy may provide a convenient and cost-effective option to mitigate the evolving pandemic and future events. In the revision, we aim to identify and characterize highly potent Nbs that are highly resistant to the variants of concern (VOCs) of SARS-CoV-2, investigate the neutralization mechanisms by structural approaches, and develop ultrapotent Nb constructs into safe and effective therapeutics. Our central hypothesis is that Nbs can be bioengineered into multivalent and ultrapotent forms to resist the mutational escape and the variants of concerns (VOCs) of SARS-CoV-2. Completion of our proposed studies will lead to cost-effective and convenient COVID-19 therapeutic candidates for translation into clinical trials. High-resolution structural studies will provide critical insights into how Nbs uniquely target the virus for high-affinity binding and neutralization. Critically, this project will serve as the testbed of our multidisciplinary platform to develop potent therapeutic and diagnostic reagents for future pandemics caused by coronaviruses or other pathogens.

项目摘要/摘要 新冠肺炎的爆发严重影响了全球健康和经济。高性价比、高度 迫切需要有效的治疗方法。骆驼VHH抗体或纳米抗体(NBS)很小，高度 稳定，易于生物工程，可以快速、经济地从微生物中制造出来。他们是 高度坚固，给药灵活，包括可能的雾化给药。把这些放在一起 NBS的独特性质使其特别适用于对抗SARS-CoV-2等呼吸道病毒 很有吸引力。 我们最近开发了一种破坏性的蛋白质组学技术，用于大规模鉴定多表位、药物- 优质国家统计局(香等人)Al，手机系统公司。2021年)。使用这项技术，我们识别了&gt；8,000个高亲和力NBS SARS-CoV-2尖峰(S)受体结合域，包括以高度中和为靶点的NBS 具有亚PM亲和力的表位，并能中和亚毫微克/毫升浓度的SARS-CoV-2，即 史无前例的抗病毒抗体片段。结构蛋白质组学揭示了多个不同的表位和 潜在的中和机制。多表位和多价结构的生物工程改进了 效价降至0.1 ng/ml以下(香等人)艾尔，科学。2020)。最近，我们展示了 超强而稳定的三聚体NB结构(PIN-21)吸入性治疗SARS的临床前疗效 新冠肺炎敏感模型中的CoV-2感染(Nabilli，et.阿尔，科学进步了。2021年)。鼻腔给药 0.6毫克/公斤的PIN-21大大减少了两个呼吸道的病毒负担。至关重要的是，PIN-21的气雾剂释放 0.2 mg/kg可通过6对数降低肺部病毒滴度，最大限度地减少感染后的肺部病理，预防病毒感染 肺炎。结合显著的稳定性和低生产成本，这种创新的疗法可能会提供一种 便利和经济高效的选择，以缓解不断演变的大流行和未来的事件。 在这项修订中，我们的目标是鉴定和鉴定高度抵抗变种的高度有效的NBS。 SARS-CoV-2的关注(VOCs)，从结构角度研究中和机制，以及 将超强的NB结构发展成安全有效的治疗方法。我们的中心假设是国家广播公司可以 被生物工程改造成多价和超强的形式来抵抗突变逃逸和变种 SARS-CoV-2的关注事项(VOCs)。 我们建议的研究的完成将导致成本效益高和方便的新冠肺炎治疗 转化为临床试验的候选人。高分辨率的结构研究将为 国家广播公司如何独特地针对病毒进行高亲和力的结合和中和。关键的是，这个项目将作为 我们多学科平台的试验台，为未来开发有效的治疗和诊断试剂 由冠状病毒或其他病原体引起的流行病。